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Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Published on: March 16, 2011

Collective peroxide detoxification determines microbial mutation rate plasticity in E. coli.

Rowan Green¹, Hejie Wang², Carol Botchey²

¹School of Natural Sciences, Faculty of Science & Engineering, University of Manchester, United Kingdom.

|July 15, 2024

Summary

Microbial mutation rates decrease with population density due to collective control of hydrogen peroxide. This density-associated mutation rate plasticity (DAMP) is linked to peroxide degradation, revealing mutation rate as a dynamic trait.

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Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

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Published on: March 16, 2011

Measuring Microbial Mutation Rates with the Fluctuation Assay

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Published on: November 28, 2019

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing

Published on: July 3, 2016

Area of Science:

Microbial genetics and evolution
Environmental microbiology
Systems biology

Background:

Mutagenesis is influenced by environmental factors, impacting evolutionary processes.
Microbial populations exhibit an inverse relationship between mutation rates and population density.
The underlying mechanism for this density-associated mutation rate plasticity (DAMP) remains largely unexplored.

Purpose of the Study:

To elucidate the mechanism driving the inverse relationship between microbial population density and mutation rates.
To investigate the role of hydrogen peroxide in mediating density-dependent mutation rate changes.

Main Methods:

Dynamical computational modeling of microbial populations.
In-culture estimation of mutation rates in Escherichia coli.
Experimental manipulation of hydrogen peroxide degradation pathways.

Main Results:

The negative correlation between mutation rate and population density is attributed to the collective regulation of hydrogen peroxide concentrations.
Escherichia coli populations lacking peroxide degradation exhibit a loss of DAMP.
Co-culturing peroxide-degradation-deficient cells with wild-type cells restores density-dependent mutation rate reduction.

Conclusions:

A mechanistic explanation for DAMP is provided, demonstrating the role of collective hydrogen peroxide control.
Mutation rate is presented as a dynamic trait influenced by microbial community composition and environmental factors.
The findings have implications across all domains of life, highlighting environmental modulation of evolutionary variation.